Home / Chondroitin B Lyase/Chondroitinase B / Recombinant P. heparinus Chondroitinase B Protein, CF

Recombinant P. heparinus Chondroitinase B Protein, CF

Catalog #: 6974-GH
4 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
6974-GH-020
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Citations (4)
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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant P. heparinus Chondroitinase B Protein, CF Summary

Product Specifications

Purity
>95%, by SDS-PAGE under reducing conditions and visualized by Colloidal Coomassie® Blue stain at 5 μg per lane
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its ability to hydrolyze chondroitin sulfate B. The specific activity is >90,000 pmol/min/μg, as measured under the described conditions.
Source
E. coli-derived p. heparinus Chondroitin B Lyase/Chondroitinase B protein
Gln26-His506, with an N-terminal Met and 6-His tag
Accession #
Q46079
N-terminal Sequence
Analysis
Inconclusive results, Met predicted. Protein identity confirmed by MS analysis of tryptic fragments.
Predicted Molecular Mass
55 kDa
SDS-PAGE
52-56 kDa, reducing conditions

Product Datasheets

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6974-GH

Product Datasheet
COA
SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

6974-GH

Formulation Supplied as a 0.2 μm filtered solution in Tris and NaCl.
Shipping The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Assay Procedure

Materials
  • Assay Buffer: 50 mM Tris, 5 mM CaCl2, pH 7.5
  • Recombinant P. heparinus Chondroitin B Lyase/Chondroitinase B (rP. heparinus Chondroitinase B) (6974‑GH)
  • Substrate: Chrondroitin Sulfate B (Sigma, Catalog # C3788), 25 mg/mL stock in deionized water
  • 96 well UV Plate (Costar, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rP. heparinus Chondroitinase B to 0.4 ng/μL in Assay Buffer.
  2. Dilute Substrate to 4 mg/mL in Assay Buffer.
  3. Load 50 μL of 0.4 ng/μL rP. heparinus Chondroitinase B into the plate, and start the reaction by adding 50 μL of 4 mg/mL Substrate. Include a Substrate Blank containing 50 μL of Assay Buffer and 50 μL of 4 mg/mL Substrate.
  4. Read at an absorbance of 232 nm in kinetic mode for 5 minutes.
  5. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

 Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol
ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

     *Adjusted for Substrate Blank 
     **Using the extinction coefficient 3800 M-1cm-1 
     ***Using the path correction 0.32 cm
     Note: the output of many spectrophotometers is in mOD. Per Well:
  • rP. heparinus Chondroitinase B: 0.020 μg
  • Chondroitin Sulfate B: 200 μg
Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Chondroitin B Lyase/Chondroitinase B

Chondroitinase B from P. heparinus is a depolymerizing lyase that is especially active on the sulfated polysaccharide dermatan sulfate (DS) (1, 2). DS is a sulfated polysaccharide that is abundantly located in the skin, but is also present in the blood vessels, heart valves, tendons, and lungs. DS may play roles in coagulation, cardiovascular disease, carcinogenesis, infection, wound repair, and fibrosis (3). DS with a repeating disaccharide unit of IdoA alpha 1-3GlcNAc is most closely related to chondroitin sulfate (CS) that has a repeating disaccharide unit of GlcA beta 1-3GlcNAc. Due to this similarity, DS was originally referred to as chondroitin sulfate B, and the DS-depolymerizing enzyme was named Chondroitinase B. DS contains more sulfate than CS at levels comparable to the anticoagulant drug heparin, which is the reason that purified commercial heparin is more likely to be contaminated by DS. Chondroitinase B can be used to specifically degrade and distinguish DS in glycosaminoglycan preparations (4, 5, 6).

References
  1. Oike, Y. et al. (1980) Biochem. J. 191:193.
  2. Prabhakar, V. et al. (2009) J. Biol. Chem. 284:974.
  3. Trowbridge, J.M. and Gallo, R.L. (2002) Glycobiology 12:117R.
  4. Gu, K. et al. (1995) Biochem. J. 312:356.
  5. Tkalec, A.L. et al. (2000) Appl. Environ. Microbiol. 66:29.
  6. Wu, Z.L. et al. (2011) Glycobiology 21:625.
Entrez Gene IDs
8251878 (P. heparinus)
Alternate Names
Chondroitin B Lyase; Chondroitinase B

Product Specific Notices

Coomassie is a registered trademark of Imperial Chemical Industries Ltd.

Citations for Recombinant P. heparinus Chondroitinase B Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

4 Citations: Showing 1 - 4
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  1. Glycosaminoglycan Domain Mapping of Cellular Chondroitin/Dermatan Sulfates
    Authors: A Persson, E Vorontsov, G Larson, J Nilsson
    Sci Rep, 2020-02-26;10(1):3506.
    Species: Human
    Sample Types: Cells
    Applications: Enzyme Activity, enzymne activity
  2. LC-MS/MS characterization of xyloside-primed glycosaminoglycans with cytotoxic properties reveals structural diversity and novel glycan modifications
    Authors: A Persson, A Gomez Tole, E Vorontsov, W Nasir, D Willén, F Noborn, U Ellervik, K Mani, J Nilsson, G Larson
    J. Biol. Chem., 2018-05-08;0(0):.
    Applications: Bioassay
  3. Xyloside-primed chondroitin sulfate/dermatan sulfate from breast carcinoma cells with a defined disaccharide composition has cytotoxic effects in vitro
    J Biol Chem, 2016-05-12;0(0):.
    Species: Human
    Sample Types: Complex Sample Type
    Applications: Bioassay
  4. Musculocontractural Ehlers-Danlos syndrome and neurocristopathies: dermatan sulfate is required for Xenopus neural crest cells to migrate and adhere to fibronectin
    Authors: N Gouignard, M Maccarana, I Strate, K von Stedin, A Malmström, EM Pera
    Dis Model Mech, 2016-04-21;0(0):.
    Species: Xenopus
    Sample Types: Protein
    Applications: Enzyme Assay

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